Unfolding and aggregation of monoclonal antibodies on cation exchange columns: effects of resin type, load buffer, and protein stability

Journal of Chromatography. a
Jing Guo, Giorgio Carta

Abstract

The chromatographic behavior of a monoclonal antibody (mAb) that exhibits a pronounced two-peak elution behavior is studied for a range of strong cation exchange resins and with varying load buffer pH and composition. Six stationary phases are considered, including two tentacle-type resins (Fractogel EMD SO3-(M) and Eshmuno S), a resin with grafted polymeric surface extenders (Nuvia S), a resin with a bimodal pore size distribution (POROS HS 50), and two macroporous resins without polymer grafts (Source 30S and UNOsphere Rapid S). The two-peak elution behavior is very pronounced for the tentacle and polymer-grafted resins and for POROS HS 50, but is essentially absent for the two macroporous resins. The extent of this behavior decreases as the buffer pH and concentration increase and, consequently, mAb binding becomes weaker. Replacing sodium with arginine as the buffer counterion, which is expected to decrease the mAb binding strength, nearly completely eliminates the two-peak behavior, while replacing sodium with tetra-n-butylammonium hydroxide, which is expected to increase the mAb binding strength, dramatically exacerbate the effect. As shown by hydrogen-deuterium exchange mass spectrometry (HX-MS), the two-peak elution beh...Continue Reading

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Citations

Feb 27, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Caleb AcquahClarence M Ongkudon
Nov 8, 2016·International Journal of Biological Macromolecules·Tsutomu ArakawaTeruo Akuta
Jan 10, 2018·Biotechnology and Bioengineering·Monika FarysRichard Kucia-Tran
Mar 18, 2020·Proceedings of the National Academy of Sciences of the United States of America·Ohnmar KhanalAbraham M Lenhoff
Jan 17, 2021·Journal of Chromatography. a·Linling Yu, Yan Sun
Aug 30, 2019·Journal of Pharmaceutical Sciences·Yoann Le BasleValérie Sautou
Apr 13, 2021·MAbs·Ohnmar Khanal, Abraham M Lenhoff

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